Abstract
The lignin modifying enzymes (LMEs) secreted by a new white rot fungus isolated from Chile were studied in this work. This fungus has been identified as a new anamorph of Bjerkandera sp. based on the sequences of the ribosomal DNA and morphological analysis at light microscopy showing hyaline hyphae without clamp connection, cylindrical conidia and lack of sexual forms, similar to those reported in other Bjerkandera anamorphs. The characterization of the culture medium for the highest LMEs production was performed in flask cultures, with a formulation of the culture medium containing high levels of glucose and peptone. The highest Mn-oxidizing peroxidase activity (1,400 U/L) was achieved on day 6 in Erlenmeyer flasks. Four peroxidases (named R1B1, R1B2, R1B3 and R1B4), have been purified by using ion-exchange and exclusion molar chromatographies. All of them showed typical activity on Mn2+ and exhibited Mn-independent activity against 2,6-dimethoxyphenol. R1B4 showed also activity on veratryl alcohol (pH 3) indicating that this enzyme belongs to the versatile peroxidase family. The high VP production capacities of this strain, as well as the enzymatic characteristics of the LMEs suggest that it may be successfully used in the degradation of recalcitrant compounds.
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References
Altschul SF, Madden TL, Schaffer AA, Zhang JH, Zhang Z, Miller W, Lipman DJ (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402
Barnett H, Hunter B (1998) Descriptions and illustrations of genera. In: Barnett H, Hunter B (eds) Illustrated genera of imperfect fungi, 4th edn. American Phytopathological Society, St. Paul, MN, pp 68–69
Böckle B, Martínez MJ, Guillén F, Martínez AT (1999) Mechanism of peroxidase inactivation in liquid cultures of the ligninolytic fungus Pleurotus pulmonarius. Appl Environ Microb 65:923–928
Bradford M (1976) A rapid and sensitive method for the quantitation of microgram quantities of proteins utilizing the principle of protein-dye binding. Anal Biochem 72:248–254
Cabana H, Jones JP, Agathos SN (2007) Elimination of endocrine disrupting chemicals using white rot fungi and their lignin modifying enzymes: a review. Eng Life Sci 7:429–456
Chivukula M, Spadaro JT, Renganathan V (1995) Lignin peroxidase-catalyzed oxidation of sulfonated azo dyes generates novel sulfophenyl hydroperoxides. Biochemistry 34:7765–7772
Davila-Vazquez G, Tinoco R, Pickard MA, Vazquez-Duhalt R (2005) Transformation of halogenated pesticides by versatile peroxidase from Bjerkandera adusta. Enzyme Microb Tech 36:223–231
Field JA, de Jong E, Costa GF, Debont JAM (1992) Biodegradation of polycyclic aromatic-hydrocarbons by new isolates of white rot fungi. Appl Environ Microb 58:2219–2226
Gardes M, Bruns TD (1993) ITS primers with enhanced specificity for basidiomycetes: application to the identification of mycorrhizae and rusts. Mol Ecol 2:113–118
Guillén F, Martínez AT, Martínez MJ, Evans CS (1994) Hydrogen peroxide producing system of Pleurotus eryngii involving the extracellular enzyme aryl alcohol oxidase. Appl Biochem Biotech 41:465–470
Günther T, Sack U, Hofrichter M, Latz M (1998) Oxidation of PAH and PAH-derivatives by fungal and plant oxidoreductases. J Basic Microb 38:113–122
Hatakka A (1994) Lignin modifying enzymes from selected white-rot fungi: production and role in lignin degradation. FEMS Microbiol Lett 13:125–135
Heinfling A, Martinez MJ, Martinez AT, Bergbauer M, Szewzyk U (1998) Purification and characterization of peroxidases from the dye-decolorizing fungus Bjerkandera adusta. FEMS Microbiol Lett 165:43–50
Kornillowicz-Kowalska T, Wrzosek M, Ginalska G, Iglik H, Bancerz R (2006) Identification and application of a new fungal strain Bjerkandera adusta R59 in decolorization of daunomycin wastes. Enzyme Microb Tech 38:583–590
Kuwahara M, Glenn JK, Morgan MA, Gold MH (1984) Separation and characterization of two extracellular H2O2 dependent oxidases from ligninolytic cultures of Phanerochaete chrysosporium. Febs Lett 169:247–250
Martínez AT (2002) Molecular biology and structure-function of lignin- degrading heme peroxidases. Enzyme Microb Tech 30:425–444
Martínez MJ, Ruiz Dueñas FJ, Guillén F, Martínez AT (1996) Purification and catalytic properties of two manganese peroxidase isoenzymes from Pleurotus eryngii. Eur J Biochem 237:424–432
Muñoz C, Guillén F, Martínez AT, Martínez MJ (1997) Laccase isoenzymes of Pleurotus eryngii: Characterization, catalytic properties, and participation in activation of molecular oxygen and Mn2+ oxidation. Appl Environ Microb 63:2166–2174
Okeke BC, Paterson A, Smith JE, WatsonCraik IA (1997) Comparative biotransformation of pentachlorophenol in soils by solid substrate cultures of Lentinula edodes. Appl Microbiol Biot 48:563–569
Palma C, Martínez AT, Lema JM, Martínez MJ (2000) Different fungal manganese-oxidizing peroxidases: a comparison between Bjerkandera sp and Phanerochaete chrysosporium. J Biotechnol 77:235–245
Romero E, Speranza M, García-Guinea J, Martínez AT, Martínez MJ (2007) An anamorph of the white-rot fungus Bjerkandera adusta capable of colonizing and degrading compact disc components. FEMS Microbiol Lett 275:122–129
Rubilar O, Feijoo G, Diez C, Lú-Chau TA, Moreira MT, Lema JM (2007) Biodegradation of pentachlorophenol in soil slurry cultures by Bjerkandera adusta and Anthracophyllum discolor. Ing Eng Chem Res 46:6744–6751
Ruiz-Duenas FJ, Morales M, Garcia E, Miki Y, Martinez MJ, Martinez AT (2009) Substrate oxidation sites in versatile peroxidase and other basidiomycete peroxidases. J Exp Bot 60:441–452
Somogyi M (1945) A new reagent for the determination of sugars. J Biol Chem 160:61–68
Tanaka T, Tonosaki T, Nose M, Tomidokoro N, Kadomura N, Fujii T, Taniguchi M (2001) Treatment of model soils contaminated with phenolic endocrine-disrupting chemicals with laccase from Trametes sp in a rotating reactor. J Biosci Bioeng 92:312–316
Tien M, Kirk TK (1988) Lignin peroxidase of Phanerochaete chrysosporium. Method Enzymol 161:238–249
Valentín L, Lú-Chau TA, López C, Feijoo G, Moreira MT, Lema JM (2007) Biodegradation of dibenzothiophene, fluoranthene, pyrene and chrysene in a soil slurry reactor by the white-rot fungus Bjerkandera sp BOS55. Process Biochem 42:641–648
Wesenberg D, Kyriakides I, Agathos SN (2003) White-rot fungi and their enzymes for the treatment of industrial dye effluents. Biotechnol Adv 22:161–187
White T, Bruns T, Lee S, Taylor J (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis M, Gelfand D, Sninsky J, White T (eds) PCR protocols: a guide to methods and applications, vol 01. Academic Press, San Diego, pp 315–322
Wirsel SGR, Leibinger W, Ernst M, Mendgen K (2001) Genetic diversity of fungi closely associated with common reed. New Phytol 149:589–598
Acknowledgments
This study has been supported by the Spanish projects S-0505/AMB0100, PGIDIT06PXIB265088PR and CTQ2007-66788, and the European project EUI2008-03703. Authors thanks M. Jurado and B. Casas by your help in ITS analysis and electrophoretic techniques. R. Taboada thanks to BES-2008-006977 PhD fellowship. T. Lu-Chau wishes to thank the Isabel Barreto program from the Galician Government for the economical support provided during the development of this work.
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An erratum to this article can be found at http://dx.doi.org/10.1007/s11274-011-0788-1
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Taboada-Puig, R., Lú-Chau, T., Moreira, M.T. et al. A new strain of Bjerkandera sp. production, purification and characterization of versatile peroxidase. World J Microbiol Biotechnol 27, 115–122 (2011). https://doi.org/10.1007/s11274-010-0435-2
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DOI: https://doi.org/10.1007/s11274-010-0435-2